• 대한암한의학회지
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• 제11권1호
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• pp.105-118
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• 2006

Two of the essential processes required for metastasis are neoangiogenesis and tumor cell invasion of basement membranes (BM) and extracellular matrix (ECM). Recently, data showed that herbs removing blood stasis has an anti-angiogenic effects. Tonifying vital Qi and eliminating pathogenic factor was a basic modality in Oriental oncology. In this study, we investigated several Qi and Blood tonics for potent angiogenic inhibitors. Methanol extracts of samples inhibited the proliferation of ECV-304 at the concentration of 100 ${\mu}g/m{\ell}$. Zizyphi Fructus, Glycyrrhizae Radix, Angelicae Gigantis Radix decreased the gelatinolytic activity of MMP-9 from ECV-304, at the concentration of 100 ${\mu}g/m{\ell}$ in gelatin zymography. In in vitro invasion assay, herbs inhibited the invasion activity of ECV-304 by 53% of control (Ginseng Radix), 39% (Zizyphi Fructus), 36% (Angelicae Gigantis Radix), 25% (Glycyrrhizae Radix). Ginseng Radix inhibited the capillary-like tube formation of ECV-304 at the concentration of 160 ${\mu}g/m{\ell}$, Angelicae Gigantis Radix and Paeoniae Radix Alba inhibited at the concentration of 320 ${\mu}g/m{\ell}$. These results indicated that Ginseng Radix, Glycyrrhizae Radix, and Angelicae Gigantis Radix could be considered as potent angiogenic inhibitiors.

• 대한화장품학회지
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• 제33권4호
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• pp.245-249
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• 2007

노화 과정 중에 일어나는 extracellular matrix (ECM)의 변성은 피부의 주름과 탄력 감소를 유발한다. 현재까지 항노화의 주요 타겟은 metalloproteases 혹은 콜라겐이나 엘라스틴 같은 구조 단백질에 집중되어 있지만, 최근 세포와 ECM 단백질(콜라겐, 피브릴린, 파이브로넥틴) 간의 상호작용이 세포의 생존과 증식, 조직의 재건에 중요한 역할을 한다고 알려졌다. 파이브로넥틴은 다른 ECM 단백질이나 인테그린 같은 세포 표면 수용체와 결합할 수 있는 부위를 가진 부착 단백질이다. 최근 보고에 따르면 세린 프로티아제들에 의해 분해된 파이프로넥틴 조각이 골아세포에서 MMPs 발현을 증가시킨다. 그러나 파이브로넥틴 조각의 사람 피부에서의 역할은 보고된 바 없다. 본 연구에서는 노인의 피부에서 파이브로넥틴 조각이 현저히 증가되어 있으며, 섬유아세포에 파이브로넥틴 조각을 처리하였을 시, MMP-1의 발현과 MMP-2의 활성이 증가한다는 것을 입증하였다. 이 결과는 파이브로넥틴 조각이 피부 노화를 유발하는 새로운 인자일 가능성을 제시하고 있다.

• Journal of Acupuncture Research
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• 제20권2호
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• pp.98-111
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• 2003

Objective : This study was designed to analyze the effects of bee venom and melittin on cell death in neuroblastoma cell line. Methods : MTT assay, morphologic method, DNA fragmentation, flow cytometry, immunocytochemistry analysis, RT-PCR and Western blot were performed. Results : The obtained results are summarized as follows: 1. The MTT assay demonstrated that neuroblastoma cell viability was significantly inhibitted dose-dependently by treatment with bee venom and melittin in comparison with control. 2. Cell culture demonstrated that control group proliferated highestly at he 5th day in comparison with the 4th day in bee venom and melittin group. And in bee venom and melitti group cell proliferation decreased 2.5 times than control group. 3. The morphologic study demonstrated that neuroblastoma cell showed apoptosis after treatment with bee venom and melittin for 6 hours using microscope. 4. The Flow cytometry demonstrated that apoptosis of neuroblastoma cell treated with bee venom and melittin was related with stop of cell cycle in stage of $G_0/G_1$. 5 .DNA fragmenation demonstrated that neuroblastoma cell treated with bee venom and melittin showed DNA ladder below 1 Kbp. 6. Immunocytochemistry assay demonstrated that Fos and MAPK which are related with cancer were down-regulated by treatment with bee venom and melittin. 7. RT-PCR analysis demonstrated that Fos and MAPK mRNA were transcripted. Fos was down-regulated form treatment with $5{\mu}g/ml$ bee venom and MAPK was down-regulated form $1{\mu}g/ml$ bee venom. 8. Western blot demonstrated that Fos was down-regulated from $1{\mu}g/ml$ bee venom whereas MAPK was expressed by $1{\mu}g/ml$ bee venom but down-regulated by $10{\mu}g/ml$ bee venom. Conclusions : We found that some cancer related genes ware down-regulated by treatment with bee venom and melittin. Further study is needed for investigating the anti-cancer effect of bee venom and melittin.

• 대한본초학회지
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• 제21권2호
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• pp.37-46
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• 2006

Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.3035-3042
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• 2015

Background: Isorhamnetin (Iso), a novel and essential monomer derived from total flavones of Hippophae rhamnoides that has long been used as a traditional Chinese medicine for angina pectoris and acute myocardial infarction, has also shown a spectrum of antitumor activity. However, little is known about the mechanisms of action Iso on cancer cells. Objectives: To investigate the effects of Iso on A549 lung cancer cells and underlying mechanisms. Materials and Methods: A549 cells were treated with $10{\sim}320{\mu}g/ml$ Iso. Their morphological and cellular characteristics were assessed by light and electronic microscopy. Growth inhibition was analyzed by MTT, clonogenic and growth curve assays. Apoptotic characteristics of cells were determined by flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay, immunocytochemistry and terminal deoxynucleotidyl transferase nick end labeling (TUNEL). Tumor models were setup by transplanting Lewis lung carcinoma cells into C57BL/6 mice, and the weights and sizes of tumors were measured. Results: Iso markedly inhibited the growth of A549 cells with induction of apoptotic changes. Iso at $20{\mu}g/ml$, could induce A549 cell apoptosis, up-regulate the expression of apoptosis genes Bax, Caspase-3 and P53, and down-regulate the expression of Bcl-2, cyclinD1 and PCNA protein. The tumors in tumor-bearing mice treated with Iso were significantly smaller than in the control group. The results of apoptosis-related genes, PCNA, cyclinD1 and other protein expression levels of transplanted Lewis cells were the same as those of A549 cells in vitro. Conclusions: Iso, a natural single compound isolated from total flavones, has antiproliferative activity against lung cancer in vitro and in vivo. Its mechanisms of action may involve apoptosis of cells induced by down-regulation of oncogenes and up-regulation of apoptotic genes.

• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9319-9325
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• 2014

Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose ($0.8{\mu}g/ml$) significantly inhibiting proliferation. The non-tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

• 대한소아치과학회지
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• 제35권4호
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• pp.597-606
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• 2008

본 연구는 TNFR family인 CD137 및 RANK, 파골세포의 CD137L와 T 세포의 RANKL 간의 역신호에 의한 이들 세포 의 역할을 알아보고자 하였다. 이에 RANKL 및 CD137L 자극으로 유도되는 역신호 전달에 의한 T 세포 활성과 파골세포분 화에 미치는 영향을 규명하고자 웅성 생쥐의 골수세포와 T 세포를 공동배양하여 다음과 같은 결과를 얻었다. 1. 생쥐 단핵세포주 및 골수유도 단핵전구세포에서 CD137L이 발현되며, CD137L 단클론 항체로 자극을 주었을 경우 파 골세포 표지단백질인 TRAP 양성 파골세포의 형성이 억제되었다. 2. 활성화된 $CD4^+$ 및 $CD8^+$ T 세포에서 RANKL을 발현하였으며 RANKL의 유사 수용체인 OPG 재조합 단백질을 처리 하여 $CD4^+$ 및 $CD8^+$ T 세포의 세포증식이 억제되었다. 이 연구의 결과는 CD137 자극에 의한 T 세포활성 및 RANK 자극에 의한 파골세포분화 및 활성이 각각 수용체에 결합하 는 라이겐드의 역신호에 의해 억제되었는데, 이는 파골세포와 T 세포의 과도한 활성을 제어하는 생체의 항상성조절에 관여하 는 기전으로 생각된다.

• 한국식품영양과학회지
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• 제41권1호
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• pp.20-25
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• 2012

본 연구는 천년초 줄기를 이용하여 항균, 항산화, 항암 등 천년초의 생리활성 효과를 연구하였으며 그 결과는 다음과 같다. 천년초 줄기의 추출물중 gram 양성균인 4가지 균주(Rhodococcus equi, Staphylococcus aureus, Clostridium perfringens, Bacillus cereus)에서 항균 효과가 나타난 것은 메탄올과 핵산층이었으며, 4가지 균주 중 Bacillus cereus를 제외한 3종에 대해서는 우수한 항균효과를 나타냈다. 천년 초 줄기의 peroxynitrite($ONOO^-$) 제거능 측정결과 핵산층을 제외한 메탄올층, 부탄올층, 수층의 항산화 활성은 천년초 줄기 추출물의 농도가 증가할수록 항산화 효과가 유의적으로(p<0.05) 높게 나타났다. 천년초 줄기 추출물의 각 용매별 DPPH 유리기 소거능은 농도가 증가함에 따라 소거능도 유의적으로(p<0.05) 증가하였으며, 부탄올>메탄올>수층>핵산층 순으로 높게 나타났다. 천년초 줄기의 암세포 증식 억제 효과에서 피부암 세포의 경우 $50\sim100{\mu}g/mL$의 농도에서 메탄올층과 핵산층이 수층과 부탄올층에 비하여 유의적으로(p<0.05) 높게 나타났다. 간암세포인 HepG2의 경우 추출물 $100{\mu}g/mL$의 농도를 기준으로 메탄올층이 유의적으로(p<0.05) 높았고, 핵산층>부탄올층>수층의 순으로 나타났다. 대장암세포인 HT-29 역시 메탄올층이 유의적으로 가장 높았고, 핵산층>수층>부탄올층 순으로 나타났다. 유방암세포인 MCF-7은 메탄올층>핵산층>수층>부탄올층 순으로 효과가 높게 나타났다. 이와 같은 결과로 볼 때 천년초줄기의 항균, 항산화, 항암 효과가 높게 나타나 줄기를 활용한 항균제품이나 여러 가지 가공제품을 개발하는데 기초 자료로 활용할 수 있다.

• BMB Reports
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• 제51권5호
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• pp.255-260
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• 2018

Wntless/GPR177 functions as WNT ligand carrier protein and activator of $WNT/{\beta}$-catenin signaling, however, its molecular role in gastric cancer (GC) has remained elusive. We investigated the role of GPR177 in gastric tumorigenesis and provided the therapeutic potential of a clinical development of anti-GPR177 monoclonal antibodies. GPR177 mRNA expression was assessed in GC transcriptome data sets (GSE15459, n = 184; GSE66229, n = 300); protein expression was assessed in independent patient tumor tissues (Yonsei TMA, n = 909). GPR177 expression were associated with unfavorable prognosis [log-rank test, GSE15459 (P = 0.00736), GSE66229 (P = 0.0142), and Yonsei TMA (P = 0.0334)] and identified as an independent risk predictor of clinical outcomes: GSE15459 [hazard ratio (HR) 1.731 (95% confidence interval; CI; 1.103-2.715), P = 0.017], GSE66229 [HR 1.54 (95% CI, 1.10-2.151), P = 0.011], and Yonsei TMA [HR 1.254 (95% CI, 1.049-1.500), P = 0.013]. Either antibody treatment or GPR177 knockdown suppressed proliferation of GC cells and sensitized cells to apoptosis. And also inhibition of GPR177 suppresses in vitro and in vivo tumorogenesis in GC cells and inhibits $WNT/{\beta}$-catenin signaling. Finally, targeting and inhibition of GPR177 with antibody suppressed tumorigenesis in PDX model. Together, these results suggest GPR177 as a novel candidate for prognostic marker as well as a promising target for treatment of GC patients.

• 동의생리병리학회지
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• 제23권5호
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• pp.1003-1011
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• 2009

This study was carried out to know the effects of Kwanjulbang-6(GJB-6) on the inhibition of arthritis. GJB-6 was orally administered to mouse with arthritis induced by collagen II . Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node and paw joint, rheumatoid factor in serum were measured in vivo. The incidence of arthritis was significantly decreased. Total cell number of draining lymph node was significantly increased compared with control. Total cell number of paw joint was significantly decreased compared with control. The absolute number of $CD19^+$, $CD8^+$, $CD3^+/CD69^+$, $CD3^+/CD49b^+$, $CD4^+/CD44^+$, $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD3^+/CD8^+$ cells in draining lymph node were significantly increased compared with control. The absolute number of $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD11b^+/Gr-1^+$ cells in paw joint were significantly decreased compared with control. The absolute number of $B220^+/CD23^+$ and $MHCII^+/CD11c^+$ cell in draining lymph node were significantly decreased compared with control. The levels of IgG was decreased and The levels of IgM was significantly decreased compared with control. Anti-collagen II in serum was significantly decreased compared with control. With the hematoxylin and eosin stain, the cartilage destruction and synovial cell proliferation were decreased compared with control. With the Masson's trichrome stain, the expression of collagen fibers was decreased compared with control. Results showed that GJB-6 had immunomodulatory effects. So we expect that GJB-6 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.